None
The invention relates to the treatment of and immunization against proliferative diseases, inflammatory diseases and pain. More specifically the invention relates to the use of metal chelating materials including, picolinic acid, fusaric acid, their derivatives, analogs and related chemicals as pharmacological and/or biological response modifier agents. It will be appreciated that the scope of the invention, as well as the scope of the appended claims, are intended to include pharmaceutically accepted salts when reference is made to the picolinic acid, fusaric acid, their derivatives, analogs and related chemicals
It will be appreciated that hereinafter the use of the term xe2x80x9cresponse modifierxe2x80x9d is intended to encompass all of the intended functions of the invention and method including antiviral, antiinfective, antiinflammatory, anticancer, antiaging, growth stimulant, vaccine and so on. Further, it will be appreciated that the broad term xe2x80x9cantiinfectivexe2x80x9d is intended to include antibacterial, antifungal, antiparasitic functions, as well as actions against any other infective agent or organism including viruses not encompassed by the term xe2x80x9cantiviralxe2x80x9d. It will also be appreciated that the term xe2x80x9cantiinflammatoryxe2x80x9d is intended to include an inflammatory response modifier, including all inflammatory responses such as production of stress proteins, white blood cell infiltration, fever, pain, swelling and so forth. Furthermore, the term xe2x80x9canalgesicxe2x80x9d is intended to include a pain reliever, whether the pain incurred is a result of disease, inflammation, trauma or psychosomatic reaction.
Researchers recently have come to appreciate the role of metal containing proteins in physiological actions and responses including cancer, pain, inflammation, proliferative and infectious diseases. Generally speaking, the inventor has studied the important function of proteins having amino acid sequences which bind metals, particularly transition metal ions therein. For example, the inventor has determined the important role zinc finger or zinc ring proteins as hormone-receptor proteins and in proliferative, inflammatory and infectious diseases. Moreover, the inventor has determined the role of other metal ion containing protein complexes, such as the role of iron finger proteins such as iron-finger hormone-receptor proteins in aging and carcinogenesis.
The inventor and others have recognized at least three efficient approaches to inhibiting zinc finger proteins: 1) disruption of the zinc finger by modification of the cysteines which are at least one of the four binding sites for Zn2+ in the zinc finger protein which results in the ejection of zinc ion; 2) removal of the zinc from the zinc finger moiety by specific chelating agents; and 3) specific chelating agents that form a ternary complex at the site of zinc binding on zinc finger proteins, resulting in inhibition of the DNA or RNA binding activity of zinc finger proteins.
Papilloma virus infection results in a number of proliferative diseases in subjects including warts induced by type 4 human papilloma virus (common warts). Moreover, papilloma virus can cause plantar ulcers as well as plantar warts. Human papilloma virus infection of the uterine cervix is the most common of all sexually transmitted diseases. Commonly known as genital warts, this wide spread virus infection is a serious disease that potentially can develop into cervical cancer. Since the virus is permanently present in cells, infection recurs in a significant percentage of patients. In many instances, conization of the uterine cervix is required to remove the infected tissue.
Condylomata acuminata, also denoted genital warts, are benign epithelial growths that occur in the genital and perianal areas and caused by a number of human papilloma viruses (HPV) including types 6, 11 and 54. These are low risk viruses which rarely progress to malignancy. However, high risk viruses such as HPV-16 and HPV-18 are associated with cervical intraepithelial cancer.
The actions of HPV are mediated by specific viral-encoded proteins which interact and/or modulate cellular DNA and proteins to produce abnormal growth and differentiation of cells. Two proteins of the HPV viral genome, E6 and E7, are well conserved among anogenital HPVs and both may contribute to the uncontrolled proliferation of basal cells characteristic of the lesions. The E7 oncoprotein is a multi-functional protein with transcriptional modulatory and cellular transforming properties. The E7 oncoprotein is denoted as a xe2x80x9czinc fingerxe2x80x9d protein because it possesses a sequence motif that is implicated in zinc binding. A strong correlation between zinc binding and the transactivation activity of E7 has been documented. The HPV-16 E6 protein is a xe2x80x9czinc fingerxe2x80x9d protein that binds DNA and may have transcriptional properties such that its function may be dependent upon the formation of zinc fingers. E6 protein can complex with the cellular tumor suppressor protein p53 and it is necessary with E7 protein for the immortalization of primary human squamous cells. Only two proteins of HPV are consistently expressed and integrated in keratinocytes, the E6 and E7 zinc finger proteins. The E6 and E7 proteins are responsible for continuous cell proliferation. About twenty HPVs are associated with ano-genital lesions and all transformed keratinocytes of these lesions contain E6 and E7 zinc finger proteins. The E6 and E7 regulate growth and transformation by interfering with cellular p53 and pRb proteins, respectively. Thus, one should be able to control or cure HPV by inactivating E6 and E7, the critical zinc finger proteins which are required for replication. When replication of the virus is halted, apoptosis of the virally-infected cells must occur. Thus, one can alter the epidemiology of, for example, carcinoma of the uterine cervix by interfering with the function of zinc finger or zinc ring proteins.
Herpes viruses, for example, Herpes Simplex Virus (HSV), has two important viral metalloproteins, a zinc finger protein and ribonucleotide reductase, an iron-containing enzyme, which are necessary for replication and propagation of the viruses. One can alter the course of herpes diseases, such as xe2x80x9cfever blistersxe2x80x9d and genital herpes, by inhibiting the two viral metalloproteins.
The human immunodeficiency virus (HIV) encodes several regulatory proteins that are not found in other retroviruses. The tat protein, which is one of these proteins, trans-activates genes that are expressed from the HIV long terminal repeat and tat is essential for viral replication. The tat protein of the HIV-1 is a zinc finger protein that when added to certain cells in tissue culture, specifically promotes growth. It has been shown that the tat protein of HIV-1 stimulates growth of cells derived from Kaposi""s sarcoma lesions of AIDS patients. Other experiments raised the possibility that tat might act as a viral growth factor to stimulate replication in latently infected cells or alter expression of cellular genes.
The nucleocapsid p7 protein of HIV has been targeted by the inventor for treatment of HIV viral infections. The p7 protein is required for the correct assembly of newly formed virus particles during the viral life cycle. Moreover, the p7 protein contains two zinc fingers that are required for the recognition and packaging of the viral RNA. Because the zinc finger domain is essential for nucleic acid binding, p7 resistant mutants are highly unlikely to occur. Thus, agents that effectively attack the two zinc finger domains of the HIV virus nucleocapsid p7 in vivo will decrease the overall number of viral particles that bud off and exit the cells to infect other cells.
The hepatitis C virus is not integrated with DNA and thus may be vulnerable to attack by specific antivirals. The hepatitis C viruses are dependent upon the Zn2+ metalloproteinases for specific viral functions. Processing at the C terminus portion of the NS2 protein of hepatitis C virus is mediated by virus encoded protease (metalloproteinases). Modification of the metalloproteinases presents an opportunity for controlling the progression of hepatitis C mediated disease.
It is of interest to note that the breast cancer susceptibility gene BRCA1 includes a zinc ring domain that is involved in protein-protein interactions or protein-DNA interactions. It also is of interest to note that the zinc ring domain of the BRCA1 has a 54% sequence similarity and 38% sequence identity with a zinc ring domain encoded by the genome of the equine herpes virus. (R. Bienstock, xe2x80x9cMolecular Modeling of Proteins Structures, Science and Medicine, January/February 1997, p.56).
From the foregoing it appears that it would be beneficial to have a product that can interfere with the formation or action of certain zinc finger proteins or zinc ring proteins to stop the progress of certain virally induced or mediated proliferative diseases or to halt the progress of viruses or malignancies dependent upon zinc finger or zinc ring proteins for their transformation and immortalization. Furthermore, it would be beneficial to provide a product that can halt the growth of other proliferative cells, such as malignant cells by chelating metal ions from zinc-dependent or iron-dependant, transition metal ion (e.g. copper, iron, etc.) dependent proteins, hormones and enzymes necessary for the replication of the malignant cells.
It is among the objects of the present invention to provide a compound which can retard the growth and proliferation of target viruses or cells by blocking the activity of metal ion-containing proteins and proteases. The present invention can retard the growth and proliferation of target infective organisms including bacteria, fungi, parasites or other infective agents by blocking the activity of metal ion-containing proteins.
The present invention also is used to retard the growth and proliferation of target viruses or cells by blocking the activity of transition metal ion-containing protein structures such as zinc finger, zinc ring proteins, iron-finger or iron-ring proteins or enzymes associated with replication.
The invention is to provide such a compound that can be added to foods, pharmaceuticals or other perishables as a preservative.
The present invention can retard the growth of premalignant and malignant cells such as virally, chemically and spontaneously transformed cells and can retard the growth of premalignant and malignant cells such as virally, chemically and spontaneously transformed cells and be administered by any acceptable route, including orally, with substantial effectiveness and minimal side effects. The invention provides a product which can be sprayed in the nostrils or inhaled to prevent or control upper respiratory diseases such as influenza, colds or pulmonary cancer.
Furthermore, the present invention is effective in a broad range of inflammatory disorders including symptoms of well-known autoimmune diseases as well as inflammatory response to infections and to chemical assault or radiation including, but not limited to, ultraviolet, atomic or medical radiation.
The present invention provides a number of preparations containing chelating agents such as picolinic acid or derivatives thereof that halts the progression of viral infections or proliferative diseases that is non-toxic to normal cells, easy to use, relatively inexpensive and well suited for its intended purposes.
According to the invention, briefly stated, a method or treatment and compound used in the method, for example, metal chelating compounds, such as picolinic acid or derivative thereof, for the treatment of infective or proliferative diseases, actinic lesions, inflammatory response, radiation assault, and cancers in human and animal subjects, as well as the method of treatment. The invention can be used orally or topically to treat or control a wide assortment of proliferative diseases or conditions, both spontaneous or induced by viruses, bacteria, fungi, chemicals and ultraviolet light for example. The metal chelating compounds bind metal, for example iron or transition metal ions such as zinc, required by enzymes, heat shock proteins or by transcription proteins found in viruses or malignant cells. By way of further example, the metal chelating compound, for example picolinic acid or its derivatives, is used to bind the zinc-containing p7 protein common to the HIV virus, thereby inactivating the virus and preventing the exit of RNA containing viruses or particles from the cells.
Moreover, the invention can be added to foods, fresh fruits, pharmaceutical agents and other perishables as preservative. The invention will exert its antifungal and antibacterial effects on the perishables, but is applied in a low, non-toxic concentration.
Another embodiment of the invention includes approximately 3 to 6 mM picolinic acid or derivative in an isotonic solution for intranasal use or inhalation to treat or prevent upper respiratory diseases.
Also, picolinic acid in 500 mg capsules given in dosages ranging from 250 mg per day to 2000 mg per day, or more, has been shown to be effective in reducing the size of tumors, such as cancerous lymph nodes and inducing necrotic tissue in the tumor.
One embodiment of a topical preparation consists of a solution of the chelator, for example, 0.01% to 99%, preferably 5% to 25%, picolinic acid in an appropriate vehicle, such as deionized water, lotion or so forth, and is applied to the lesion one or two times a day. The preparation can be applied to skin to control acne, warts and herpes infections and to toe nails and finger nails, for example, to treat fungal infections. In another embodiment, the topical preparation consists of an ointment or cream containing approximately 0.5% to 99%, preferably 5% to 20% picolinic acid which is applied once or twice daily to the lesion and to a bandage placed on the lesion. The ointment or cream can be instilled intravaginally to retard sexually transmitted viral diseases.
The various embodiments of the topical preparation can be used to treat papilloma and herpes viral diseases and to retard the papilloma, herpes and HIV 1 viruses as well as proliferative diseases such as psoriasis, actinic lesions and skin cancer.
Various active derivatives that maintain their activity and stability when after systemic administration are provided. Slow release oral formulations can be used to treat diseases for the digestive tract. The active derivatives can be administered orally, parenterally, by inhalation, transdermally or by any other appropriate method to control proliferative diseases, cancers, viral infections, HIV, and any other condition wherein the causative agent includes a zinc-containing protein, whether the zinc-containing protein is a zinc finger protein, a zinc ring protein, or other type of zinc or metal containing structure heretofore unidentified or undetected, wherein the metal containing segment is required for protein stability and configuration.
It will be appreciated that other appropriate chelating materials such as the derivative of picolinic acid or fusaric acid or even unrelated compounds also may be used. It also will be appreciated that, although 5% to 20% topical preparations of the picolinic acid are described, a broader range of concentrations may be used. For example from approximately 0.001% to 99.9% metal chelating agent may be used. Further, the systemic doses may be altered or adjusted to ranges greater or lesser than those described, depending on toxicity and patient response, without departing from the scope of the appended claims.